Note: Descriptions are shown in the official language in which they were submitted.
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ANTI-WART PHARMACEUTICAL COMPOSITION AND METHOD FOR
TREATING WART
BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to an anti-wart
pharmaceutical composition containing a water-soluble
extract from a plant of Solanum genus and a method for
treating wart using the water-soluble extract.
2. Description of the Related Art
Wart is a kind of dermatosis that occurs on the skin
or mucous of human or animal subjects, and is a benign
epidermal hyperplasia caused by infection of Human
papilloma virus (HPVs). In general, based on the site
where the wart forms, wart can be divided into cutaneous
wart and mucosal wart. The epidermal cells of skin or
mucosa affected by HPVs are subjected to benign
proliferation and hyperkeratosis, thereby resulting in
formation of papules or nodules with roughened surfaces
on the skin or mucosa.
In clinical diagnosis, wart is divided into four
types according to symptoms thereof: (1) common wart
(Verruca vulgaris), scaly and roughened papule(s) or
nodule(s) whichusually occur on hands or fingers; (2)
flat wart (Verruca pdana juvenilis), slightly raised and
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flat-topped papule(s) which usually occurs on faces,
hands, and lower legs; (3) plantar wart (Verruca
plantaris), a thick and hyperkeratotic papule which
usually occurs at pressure points of soles and toes and
causes painful feeling for a patient when the papule is
pressed; and (4) genital wart (Condyloma acuminatum), a
cauliflower-like lump formed on skin or mucosa of
genitalia and anus, which is a highly infectious and
sexually transmitted disease (STD).
Nowadays, warts are treated using the following
remedies based on the type, size, number, and lesion site
thereof:
(1) Topical therapy, e.g., podophyllin (Trade name:
Wartec ) and imiquimod (trade name: Aldara ) for treating
genital wart, 5-fluorouracil for treating genital wart
and cutaneous wart, and bleomycin (Trade name: Bleocin )
for treating plantar wart and warts formed on palm;
(2) Laser treatment, e.g., pulsed dye laser and carbon
dioxide laser, especially for treating plantar wart and
warts formed on palm; and
(3) Cryotherapy using liquid nitrogen and suitable
for treating most warts, less effective for warts formed
on palm, sole, and toe.
Each of the aforesaid remedies is suitable only for
a specific type of wart, rather than all types of warts.
Moreover, in clinical diagnosis, there are inferior
effect problem and side effect drawback for the aforesaid
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remedies. For example, topical therapy used for a long
period of time will cause recurrence of warts and treating
effect thereof is unsatisfactory. Laser treatment and
cryotherapy will cause severe side effects, such as pain,
ulceration, and scarring. Some patients, especially
children, are likely to refuse treatment due to the
unendurable severe side effects.
To overcome the aforesaid drawbacks, researches have
focused on finding active components from traditional
Chinese medicines or herbs to develop a drug that is
effective for treating warts and that has no undesired
side effects.
It is known that Solanum incanum L. (also known as
Solanum incanum Ruiz. & Pay., Solanum undatum, Solanum
coagulans Forsskal in Latin, and bitter apple in English)
contains steroidal glycoalkaloid which exhibits
anti-cancer activity. In addition, many plants of the
Solanum genus are reported to contain steroidal
glycoalkaloid, including, for example, Solanum indicum,
Solanum nigrum, also known as Lohg Kui in Chinese and black
nightshade in English, Solanum capsicastrum (known as
false Jerusalem cherry in English) , Solanum xanthocarpum,
Solanum melongena, Solanum coagulans, Solanum tuberosum,
Solanum sodomeum, Solanum turburosum, Solanum
aculeastrum, Solanum lycocarpum, Solanum khasianum,
Solanum suaveolens, Solanum uporo, Solanum abutiloides,
Solanum coccineum, Solanum unguiculatum, Solanum
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robustum, Solanum anguivi, Solanum platanifolium,
Solanum mammosum, etc. The common steroidal
glycoalkaloids contained in plants of Solanum genus
include solamargine and solasonine.
US 7,078,063 B2 issued to the inventors of this
invention discloses a water soluble extract from a plant
of Solanum genus, especially Solanum incanum L., and a
process for preparing the same. The water soluble
extract consists essentially of at least 60% of
solamargine and solasonine.
In the aforesaid US patent, the inventors found that
the water soluble extract exhibits inhibition of growth
of tumor/cancer cells, in particular liver tumor cells,
lung cancer cells, and breast cancer cells. In this
invention, the inventors unexpectedly found that the
water soluble extract is effective in treating warts.
SUMMARY OF THE INVENTION
Therefore, according to a first aspect, this
invention provides an anti-wart pharmaceutical
composition which includes a water-soluble extract from
a plant of Solanum genus, the water-soluble extract
comprising at least 60% of solamargine and solasonine.
In a second aspect, this invention provides a method
for treating wart, which comprises applying to a subject
in need of such treatment a water-soluble extract from a
plant of Solanum genus, the water-soluble extract
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comprising solamargine and solasonine.
Ina third aspect, this invention relates to the use
of a water-soluble extract from a plant of Solanum genus
in the manufacture of a medicament for treating wart, the
5 water-soluble extract comprising solamargine and
solasonine.
BRIEF DESCRIPTION OF THE DRAWINGS
The above and other objects, features and advantages
of this invention will become apparent with reference to
the following detailed description and the preferred
embodiments taken in conjunction with the accompanying
drawings, in which:
Fig. 1 shows photos illustrating that lesions
(indicated by arrows) of test subject C are gradually
ameliorated by treating the same with a Solanum incanum
L. gel;
Fig. 2 shows photos illustrating that lesions
(indicated by arrows) of test subject D are gradually
ameliorated by treating the same with a Solanum incanum
L. gel;
Fig. 3 shows photos illustrating that lesions
(indicated by arrows) of test subject E are gradually
ameliorated by treating the same with a Solanum incanum
L. gel; and
Fig. 4 shows photos illustrating that lesions
(indicated by arrows) of test subject F are gradually
ameliorated by treating the same with a Solanum incanum
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L. gel.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS
It is to be understood that, if any prior art
publication is referred to herein, such reference does
not constitute an admission that the publication forms
a part of the common general knowledge in the art, in
Taiwan or any other country.
For the purpose of this specification, it will be
clearly understood that the word "comprising" means
"including but not limited to", and that the words
"comprises," "contain" and variants thereof have a
corresponding meaning.
Unless otherwise defined, all technical and
scientific terms used herein have the meaning commonly
understood by a person skilled in the art to which this
invention belongs. One skilled in the art will recognize
many methods and materials similar or equivalent to those
described herein, which could be used in the practice of
this invention. Indeed, this invention is in no way
limited to the methods and materials described. For
clarity, the following definitions are used herein.
The present invention provides an anti-wart
pharmaceutical composition, which includes a
water-soluble extract from a plant of Solanum genus. The
water-soluble extract comprises solamargine and
solasonine. Preferably, the water-soluble extract
comprises at least 60 wt% of solamargine and solasonine,
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more preferably, 60 wt%-90 wt% of solamargine and
solasonine.
The process for preparing the water-soluble extract
has been disclosed in US 7,078,063 B2 and includes the
steps of:
(a) subjecting a plant material of a plant of Solanum
genus to an extraction treatment using an acidic
aqueous solution with a pH value of 3--5, such that
an aqueous solution is obtained;
(b) adjusting the pH value of the aqueous solution
obtained in step (a) to pH 8--10 with a base, such
that a precipitate is formed;
(c) washing the precipitate formed in step (b) with
water, followed by drying, such that a dried product
is obtained;
(d)admixing the dried product obtained in step (c)
with chloroform, followed by addition of a suitable
amount of a 100% alcohol, such that a
chloroform-alcohol mixture is formed;
(e) mixing the chloroform-alcohol mixture formed in
step (d) with a water/alcohol solution having a
predetermined water:alcohol ratio, such that a
mixture containing a chloroform-based layer and a
non-chloroform-based layer is obtained;
(f)removing the chloroform-based layer from the
mixture obtained in step (e), followed by addition
of a suitable amount of water; and
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(g) obtaining a supernatant from the resultant mixture
of step (f) , followed by drying the supernatant,
wherein the resultant dried product is able to be
directly dissolved in water to form a yellowish
clear and transparent aqueous solution.
Preferably, in step (a) , the plant material of the
plant of Solanum genus has been chopped in a preliminary
treatment.
Preferably, in step (a) , the plant material is at
least one of the fruit, root, stem, and leaf of the plant
of Solanum genus. In a preferred embodiment of this
invention, the plant material used in step (a) is the fruit
of the plant of Solanum genus.
The inventors found that certain factors might affect
the content and proportion of solasonine and solamargine
in the water-soluble extract obtained using the aforesaid
process. These factors include the species of the plant
of Solanum genus and the part/parts of the plant used in
the extracting process, as well as the types of alcohol
and base used. Therefore, a skilled artisan can prepare
a desired water-soluble extract by selecting a suitable
species of the plant of Solanum genus and using a suitable
part or parts of the plant, in conjunction with
appropriate operating conditions.
Preferably, the water-soluble extract is obtained
from a plant of Solanum genus selected from the group
consisting of Solanum incanum L., Solanum indicum,
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Solanum nigrum, Solanum capsicastrum, Solanum
xanthocarpum, Solanum melongena, Solanum coagulans,
Solanum tunigrum, Solanumsodomeum, Solanum turburosum,
Solanum aculeastrum, Solanum lycocarpum, Solanum
khasianum, Solanum suaveolens, Solanum uporo, Solanum
abutiloides, Solanum coccineum, Solanum unguiculatum,
Solanum robustum, Solanumanguivi, Solanumplatanifolium,
and Solanum mammosum. In a preferred embodiment of this
invention, the water-soluble extract is obtained from
Solanum incanum L..
In preliminary human tests done by the inventors, the
results show that the water-soluble extract is effective
in treating or alleviating warts including cutaneous
warts and mucosal warts without undesired side effects,
e.g., ulceration or erythema. Therefore, the present
invention provides a method for treating wart, which
comprises applying to a subject in need of such treatment
the water-soluble extract.
The wart that can be treated by the anti-wart
pharmaceutical composition of this invention includes
common wart, flat wart, plantar wart, and genital wart.
The anti-wart pharmaceutical composition of this
invention can be administered via e.g., parenteral,
transmucosal, transdermal, or topical route.
The parenteral route includes subcutaneous injection,
intraepidermal injection, intradermal injection, and
intralesional injection.
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For the parenteral route, the anti-wart
pharmaceutical composition of this invention can be
formulated into an injection product using techniques
known to a skilled artisan, e.g., sterile aqueous solution
5 or dispersion.
In this invention, the injection product can be
prepared by mixing the water-soluble extract with a
pharmaceutically acceptable carrier that is widely
employed in drug-manufacturing technology.
10 The
pharmaceutically acceptable carrier comprises
one or more reagents, including, for example, water,
saline, buffer solution (Phosphate Buffered Saline (PBS),
Ringer's solution or Hank's solution), emulsifier,
suspending agent, decomposer, pH adjusting agents,
stabilizing agent, chelating agent, preservative,
diluents, absorption delaying agent, liposome, etc. The
choice and amount of these pharmaceutically acceptable
carriers are within the expertise of those skilled in the
art.
Moreover, the anti-wart pharmaceutical composition
of this invention can be administered by topical routes,
e.g., dermal, buccal, genital, vaginal, anal, and rectal
routes. In an embodiment of this invention, the anti-wart
pharmaceutical composition of this invention is
administered by dermal route. In another embodiment of
this invention, the anti-wart pharmaceutical composition
of this invention is administered by genital route.
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The pharmaceutical composition according to this
invention can be formulated into a suitable dosage form
for topical administration using technology well known
to those skilled in the art, which includes, but is not
limited to, external preparations, effervescent tablets,
suppositories, and the like.
In a preferred embodiment of this invention, the
anti-wart pharmaceutical composition is formulated into
an external preparation by admixing the water-soluble
extract according to this invention with a base that is
well known and commonly used in the art.
In this invention, the external preparation includes,
but is not limited to, emulsion, gel, ointment, cream,
patch, liniment, powder, aerosol, spray, lotion, serum,
paste, foam, drop, suspension, and tincture. In an
embodiment of this invention, the anti-wart
pharmaceutical composition is in the form of gel.
For example, the base suitable for producing the
external preparation according to this invention may
include one or more of the following additives: water,
alcohols, glycol, hydrocarbons (e.g., petroleumjellyand
white petrolatum), wax (e.g., paraffin and yellow wax),
preserving agents, antioxidant, surfactants, absorption
enhancers, stabilizing agents, gelling agents (e.g.,
carbopoll'974P, microcrystalline cellulose, and
carboxymethyl cellulose), active agents, humectants,
odor absorbers, fragrances, pH adjusting agents,
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chelating agent, emulsifier, occlusive agents,
emollients, thickeners, solubilizing agents,
penetration enhancers, anti-irritants, colorants, and
propellants. The choice and amount of these additives are
within the expertise of those skilled in the art.
This invention also provides a method for treating
wart, which comprises applying to a subject in need of
such treatment the water-soluble extract or the anti-wart
pharmaceutical composition.
The dosage and the frequency of administration of the
anti-wart pharmaceutical composition according to this
invention may vary depending on the following factors:
the severity of the disease to be treated, the route of
administration, and the weight, age, physical condition
and response of the subject to be treated. For instance,
the daily dosage of the pharmaceutical composition
according to this invention may be 10 to 20 mg/cm2 of the
lesion area and one to six times per day.
This invention will be further described by way of
the following examples. However, it should be understood
that the following examples are solely intended for the
purpose of illustration and should not be construed as
limiting the invention in practice.
Examples
Example 1: Preparation of the water-soluble extract
The water-soluble extract was prepared based on the
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procedure as disclosed in Example 1 of US 7,078,063 B2.
Specifically, 500 g of ripe fruit of Solanum incanum L.
was ground subsequent to addition of 1000 ml pure water.
To the resultant aqueous mixture, 99.5% of acetic acid
was added dropwise to adjust the pH value to 4.0, followed
by shaking at room temperature for 12 hrs. A supernatant
was obtained by centrifuging the aqueous mixture at 4500
rpm for 12 hours, and 33% NH4OH basic solution was added
thereto dropwise to adjust the pH value of the supernatant
to 9.0, and a precipitate was formed. The precipitate was
obtained by conducting centrifugation at 4,500 rpm for
10 minutes(Beckman Coulter, Avanti J-25, JA-14 Rotor),
and the residual basic solution present therein was
removed by washing the precipitate with water, followed
by centrifugation at 4,500 rpm. The precipitate thus
obtained was suspended in distilled water and subjected
to lyophilization (Virtis, Freezemobile 12ES) to obtain
5 g of dried powder.
2 g of the dried powder was dissolved in 50 ml
chloroform in reagent grade, followed by addition of 40
ml of 100% methanol and shaking to form a uniform
suspension. A supernatant was obtained by centrifugation
at 4,500 rpm. 70 ml of methanol:water solution (1:1) was
added to the supernatant and mixed well. The mixture
obtained was centrifuged at 12,000 rpm for 10 min. The
resultant supernatant was taken out, and 120 ml distilled
water was added thereto and shaken well. The supernatant
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was further centrifuged at 12,000 rpm for 10 min so as
to remove the precipitate. The resultant supernatant was
subjected to concentration under reduced pressure at 55 C
to remove methanol, followed by lyophilization to obtain
dried powder of the water-soluble extract.
Example 2: Therapeutic effect and safety of the
water-soluble extract in treating human cutaneous wart
and mucosal wart
In order to examine the therapeutic effect and safety
of the water-soluble extract as prepared in the above
Example 1 in treating human cutaneous wart and mucosal
=
wart, the following tests were conducted.
A. Preparation of gel containing the water-soluble
extract
4 g of carbopol 974P, which was used as a gelling agent
and is commercially available from Lubrizol Advanced
Materials, Inc., KY 40258, USA, was dissolved in 50 g pure
water, followed by sequential addition of 30 g of
propylene glycol and 7 g of the dried powder of the
water-soluble extract obtained in Example 1 and mixing
well. The mixture was heated in a heating vessel at a
temperature of 50 C to 60 C for 20 minutes, followed by
cooling at room temperature. The cooled mixture was added
with triethanolamine to adjust pH to 7.0 0.5.
Subsequently, water was added until the total weight of
the mixture reached 100 g, thereby obtaining a gel
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containing 7% (w/w) of the water-soluble extract
(hereinafter referred to as Solanum incanum L. gel).
B. Test subjects and clinical information thereof
5 Six test subjects
(test subjects A to F) participating
in the following tests were enrolled from the outpatient
department of National Cheng Kung University Hospital,
Taiwan. The test subjects A and B suffered from mucosal
wart, and the test subjects C to F suffered from cutaneous
10 wart. The clinical
information of the six test subjects,
including gender, age, wart type, and lesion location,
are outlined in Table 1.
Table 1
Test Age Wart type Lesion
location
Gender
subject (y/)
A 32 Female Mucosal
wart External genital
21 Female Mucosal wart External
genital
28 Female Cutaneous wart Finger tip
21 Male Cutaneous wart Finger tip, finger
26 Male Cutaneous wart Finger
33 Male Cutaneous wart Finger tip, palm
C. Pre-clinical test
Prior to the start of the test (i.e., on Week 0),
selected lesions on each of the enrolled test subjects
were photographed. The Solanum incanum L. gel prepared
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in section A of Example 2 was applied to the selected
lesions and normal skin areas that surround the selected
lesions. Each of the normal skin areas had inner and outer
peripheries and the distance between the inner and outer
peripheries of each of the normal skin areas was not
greater than 0.5 cm. The treated dosage was 10-20 mg/cm2,
and twice a day. The selected lesions of each of the test
subjects were photographed at predetermined evaluation
times (i.e., at the ends of weeks 1, 2, 3, 4, 7, 8, 13,
20 and 29, see Table 2) , and evaluations for therapeutic
effect and safety of the Solanum incanum L. gel were
conducted according to the procedures set forth in
sections D and E below. The test was finished when the
lesions of each of the test subjects were completely cured
and the outer appearance of the lesion skin was recovered
to a normal state.
Table 2
Test Evaluation time (week)
subject Oa lb 2 3 4 7 8 13 20 29
A +c + d
+ + - - -
+ - - + -
+ - + - -
+ - + - +
+ - - + _ e f
a:"0" indicates week 0, i.e., prior to
application of the Solanum incanum L. gel
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b: "1" means week 1 after application of the
Solanum incanum L. gel
C: "+" means evaluation was conducted
d:"-"means no evaluation was conducted
e: "(0;1" means evaluation was conducted only on
the lesions at finger tip
f: ""means evaluation was conducted only on the
lesion on palm
D. Evaluation of therapeutic effect
The photos of the selected lesions of the test
subjects as obtained at Week 0 (i.e., prior to the
application of the Solanum incanum L. gel) and as obtained
at the evaluation times were compared by a dermatologist
to determine the improvement of lesions treated with the
Solanum incanum L. gel.
E. Evaluation of safety
Evaluation of safety was conducted by a researcher
at the predetermined evaluation times. The test subjects
were interrogated by the researcher in respect to the
appearance of any adverse skin response (including
ulceration, irritation and erythema).
Results:
A. Evaluation of the therapeutic effects of the Solanum
incanum L. gel
Compared with the photos of the selected lesions of
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of each of the test subjects C to F treated with the
Solanum incanum L. gel greatly improved with time. After
the test was finished, the test subjects C to F were
further subjected to the follow-up study for six months
to two years to determine whether the lesion area that
had already been cured by the Solanum incanum L. gel was
recurrent. Based on the results of the follow-up study,
no warts were found to have recurred for all of the test
subjects (data not shown).
The aforesaid results show that the Solanum incanum
L. gel of this invention could be effectively used to
treat cutaneous wart and mucosal wart.
B. Evaluation of the safety of the gel containing the
water-soluble extract
In the safety test, only a few of the test subjects
experienced temporary and mild adverse skin responses,
including irritation or erythema, which, however, did
not halt them from completing the test (data not shown).
In view of the aforesaid, the inventors contemplate
that the water-soluble extract from a plant of Solanum
genus according to this invention can be developed into
an anti-wart drug for long-term use in treating warts
without adverse side effects.
